ANTI-VIRAL GUANOSINE-RICH OLIGONUCLEOTIDES

Abstract: A method and composition for treating viral infection in vitro and in vivo using guanosine-rich oligonucleotides. The oligonucleotides have sufficient guanosine to form a guanosine tetrad. They can be composed of at least about 40 % guanosine nucleotides. Also provided are oligonucleotides of at least two runs of at least two guanosines. Also provided are guanosine-rich oligonucleotides and methods for treating viral infections in humans, and a method for designing guanosine-rich oligonucleotides having anti-viral activity.

Description: ANTI-VIRAL GUANOSINE-RICH OUGONUCLEOTIDES BACKGROUND OF THE INVENTION Field of the Invention The present invention relates generally to the field of oligonucleotide chemistry and anti-viral pharmacotherapy. More specifically, the present invention relates to novel guanosine-rich oligonucleotides and their use as novel anti- viral agents. Description of the Related Art Previously, it was believed that "antisense" oligonucleotides inhibit viruses by interfering with protein translation via an RNA:DNA duplex structure. More recent research, however, indicates a variety of possible mechanisms by which oligonucleotides inhibit viral infections. For example, oHgodeoxycytidine (poly SdC) inhibits HIV-1. Marshall et al., PNAS (1992) 89:6265-6269, discussed the potential mechanism (competitive inhibition) by which oHgodeoxycytidine directly inhibits viral reverse transcriptase. Poly SdC also inhibited AMV reverse transcriptase and Pol I (Klenow fragment) and polymerase α, β and γ. Previously, Matsukura et al., PNAS (1987) 84:7706-7710, used a similar phosphorothioate derivative of oHgodeoxycytidine to demonstrate inhibition of HIV-1 in culture. Marshall and Caruthers, Science (1993) 259:1564-1569, reported the use of diphosphorothioate oligonucleotides, e.g., antisense specific, random nucleotide combinations and oHgodeoxycytidine against HIV-1. In all cases, the mechanism of action was attributed to a direct inhibition of HIV- 1 reverse transcriptase. Other potential mechanisms of anti- viral action of oligonucleotides were postulated by Boiziau et al., PNAS (1992) 89:768-772, e.g., promotion of RNAse H activity and inhibition of reverse transcriptase initiating cDNA synthesis. In addition, Goa et al., Molecular Pharmacology (1992) 41:223-229 reported that phosphorothioate oligonucleotides inhibit human DNA polymerases and RNAse H, and the adsorption or penetration of the virus into cells. Iyer et al., Nucleic Acids Research (1990) 18:2855-2859 report that if a base was removed from an anti-sense polynucleotide forming an abasic site, the compound did not lose its activity which argues against the need for the formation of an RNA:DNA antisense mediated hybrid for anti- viral activity. Stein et al. have characterized the interaction of poly SdC with the V3 loop of HIV-1 gpl20, and postulated that the specific interaction of poly SdC with the HIV-1 V3 loop may be a mechanism by which an oligonucleotide could inhibit HIV-1 in vivo. It is known that synthetic oligonucleotides may be designed which are capable of binding to duplex DNA to form triplex DNA. See U.S. Patent No. 5,176,996 Hogan & Kessler issued January 5, 1993. This application describes a method for making synthetic guanosine-rich oligonucleotides which are targeted to specific sequences in duplex DNA and which form colinear triplexes by binding to the major groove of the DNA duplex. SUMMARY OF THE INVENTION In one embodiment of the present invention, there is provided a method of treating a pathophysiological state caused by a virus, comprising the step of administering a pharmacological dose of an oligonucleotide, said dose being sufficient to inhibit production of said virus, wherein said oligonucleotide contains a high percentage of guanosine bases. In a particular embodiment, the oligonucleotide has a three dimensional structure and this structure is stabilized by guanosine tetrads. In a further embodiment, the oligonucleotide has two or more runs of two contiguous deoxy guanosines. In another embodiment of the present invention, there is provided a method of treating a pathophysiological state caused by a herpes simplex virus, comprising the step of administering a pharmacological dose of an oligonucleotide, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains a high percentage of guanosine bases. In a particular embodiment, the oligonucleotide has a three dimensional structure and this structure is stabilized by guanosine tetrads. In a further embodiment, the oligonucleotide has two or more runs of two contiguous deoxyguanosines. In yet another embodiment of the present invention, there is provided a method of treating a pathophysiological state caused by human immunodeficiency viruses, comprising the step of administering a pharmacological dose of an oligonucleotide, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains a high percentage of guanosine bases. In a particular embodiment, the oligonucleotide has a three dimensional structure and this structure is stabilized by guanosine tetrads. In a further embodiment, the oligonucleotide has two or more runs of two contiguous deoxyguanosines. In still yet another embodiment of the present invention, there is provided a method of treating a pathophysiological state caused by human papilloma virus, comprising the step of administering a pharmacological dose of an oligonucleotide, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains a high percentage of guanosine bases. In a particular embodiment, the oligonucleotide has a three dimensional structure and this structure is stabilized by guanosine tetrads. In a further embodiment, the oligonucleotide has two or more runs of two contiguous deoxyguanosines. In still yet another embodiment of the present invention, there is provided a method of treating a pathophysiological state caused by human cytomegalovirus, comprising the step of administering a pharmacological dose of an oligonucleotide, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains a high percentage of guanosine bases. In a particular embodiment, the oligonucleotide has a three dimensional structure and this structure is stabilized by guanosine tetrads. In a further embodiment, the oligonucleotide has two or more runs of two contiguous deoxyguanosines. In still yet another embodiment of the present invention, there is provided a method of treating a pathophysiological state caused by adenovirus, comprising the step of administering a pharmacological dose of a oligonucleotide, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains a high percentage of guanosine bases. In a particular embodiment, the oligonucleotide has a three dimensional structure and this structure is stabilized by guanosine tetrads. In a further embodiment, the oligonucleotide has two or more runs of two contiguous deoxyguanosines. In still yet another embodiment of the present invention, there is provided a method of treating a pathophysiological state caused by hepatitis B virus, comprising the step of administering a pharmacological dose of a oligonucleotide, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains a high percentage of guanosine bases. In a particular embodiment, the oligonucleotide has a three dimensional structure and this structure is stabilized by guanosine tetrads. In a further embodiment, the oligonucleotide has two or more runs of two contiguous deoxyguanosines. In still yet another embodiment of the present invention, there is provided a guanosine-rich oligonucleotide having a three dimensional structure, wherein the three dimensional structure is stabilized by guanosine tetrads or at least two runs of two contiguous deoxyguanosines and wherein these oligonucleotides exhibit anti-viral activity. In a further embodiment, the oligonucleotides of the present invention have partially or fully phosphorothioated internucleoside linkages (backbones) or other chemical modifications. In a further embodiment, the oligonucleotides of the present invention have chemically modified or unnatural (synthetic) bases. BRIEF DESCRIPTION OF THE DRAWINGS Figure IA shows a 1973 base pair Hind III to Eco Rl sub fragment of the Friend Murine Leukemia Virus (FMLV) clone 57 genome. Figure IB shows a 172 base pair (Hind III to Stu I) fragment which is an expanded portion of the 1973 base pair fragment. Within this fragment is the purine rich target to which triple helix forming oHgonucleotides are directed. Figure 1C shows the entire Hind III/Eco Rl FMLV fragment cloned into the pT7-2 plasmid (United States Biochemical Corporation) yielding p275A. In this recombinant the Hind III site is 10 base pairs downstream of the T7 mRNA start site. The 5' portion of the triple helix target region is 63 base pairs downstream of the mRNA start and the Dde I site is 131 base pairs downstream of the mRNA start site. Figure ID shows the Hind III/Eco Rl FMLV fragment was cloned into pBS (Stratagene) yielding pBSFMLV. The Hind III site, triple helix target site and Dde I site are respectively 50, 103 and 171 base pairs downstream from the mRNA start site. Figure 2 shows that G-Rich phosphorothioated-oligonucleotides induced reduction in HSV-2 viral titer. VERO cells infected with HSV-2 were treated with various concentrations of the indicated drug. The results are plotted as percent virus yield relative to VERO cells infected with virus but not treated with drug (titer = 1). The filled square (B 106-62) represents a single concentration point (20 μM) for this oligonucleotide. B 106-96 is the fully phosphorothioated version of B106-62. B106-97 is the fully phosphorothioated version of B106-71. ACV (4a and 4b) is acyclovir tested against two different stock concentrations of HSV-2 strain HG52. In two experiments, after virus infection and before reapplication of oligonucleotide, the cells were rinsed with a pH 3 buffer in order to remove all virus not yet internalized (96p3 and 97p3). Figure 3 shows that the MT-2 cells infected with 0.01 m.o.i. of HIV-1 were treated with various concentrations of oligonucleotide or AZT or ddC. The data represents the number of viable cells remaining in the culture dish, i.e., not undergoing virus induced cytopathic effects (CPE). In this graph, 100% is the level of CPE occurring in cultures infected with virus but not treated with any drug. Figure 4 shows the culture media taken from NIH3T3 cells chronically infected with FMLV was mixed with various concentrations of 1100-51 or 1100-12 (fully phosphorothioate version of 1100-00). The mixtures were then assayed for the presence of viral reverse transcriptase. The data is presented as a percent of measurable reverse transcriptase in culture medium not treated with oligonucleotide. Figures 5A, 5B and 5C show the radio-labelled ( 32 P) full-length or truncated mRNA transcripts were analyzed by polyacrylamide gel electrophoresis, and then quantitated by cutting out the specific transcript and measuring the radioactivity in a scintillation counter. Figure 5A shows that the reduction in full length transcripts directed by the T7 and T3 promoter when 1100-51 (anti-parallel triple helix forming oligonucleotide; FMLV2ap) was added. Samples in which no oligonucleotide was added were counted and used as 100% transcription reference points. In all other reactions 4 x lO^M of G101-50 (4e-6) was added and where indicated G101-50 plus 1100-51 at concentrations ranging from 2 x 10 '9 to 2 x 10 "6 M (2e-9 to 2e-6). Figure 5B shows the reduction in full length transcript by 1100-01 (FMLV2p). T7 directed transcripts were treated as in Figure A. G101-50 was added to each reaction except the control (no oligo) with or without various concentration of 1100-01 or 1100-11 (26% G-ctl). Figure 5C shows the analysis of truncated (63 base pair) transcript. Figure 6 shows inhibition of HIV-1 induced syncytia formation four days post-infection. SUP Tl cells were infected with H_V-1 DV for four hours and then treated with various concentrations of oligonucleotides. Four days post-infection cells were scored for syncytia formation. All assays were performed in quadruplicate and the average values used to plot this graph. The legend to the right of the graph indicates the symbol used for each oligonucleotide tested. Figure 7 shows continued suppression of HIV-1 p24 production seven days post removal of oligonucleotide. Four days post-infection with HIV-1 DV , the media from infected cells treated with oligonucleotides (2.5 μM) was removed and replaced with fresh media without oligonucleotide. The presence of viral p24 antigen was then assayed 7 and 11-days post infection. All samples were assayed in quadruplicate and the average values used to plot this graph. The legend to the right of the graph indicates the symbol used for each oligonucleotide tested. Figure 8 shows a Dixon Plot of random oligonucleotide 1232 (SEQ. ID. NO. 41) obtained from kinetic analysis of inhibition of HIV-RT with respect to dNTP. The inhibition constant Kj was determined by simultaneously varying dNTP (without dATP) concentrations at the same time as inhibitor (oligonucleotide 1232). The determination was performed at 0.125 mM, 0.25 mM and 0.5 mM dNTP concentrations with constant Primer-Template concentration of 0.2 pM. HIV-RT was used at 1 unit in each reaction. The reported values are the result of simultaneous independent duplicates determinations. Figure 9A reveals PBMCs derived from HIV-1 positive patients were mixed with HIV-1 negative PBMCs in culture medium containing drug. On day 7 the cocultures were washed and resuspended in fresh medium containing drug. The p24 levels in medium collected on day 7 (before medium change) and day 10 were assayed for p24. Figure 5B HIV-1 negative PBMCs from two different donors were infected with HIV-1 DV and then incubated in the presence of drug for 10 days at which time the culture medium was assayed for the presence of p24 antigen. Figures 10A and 10B show inhibition of binding of V3 loop specific Mabs to HIV-1 gpl20 by phosphorothioate containing oligonucleotides. Matched sequence oligonucleotides with either phosphodiester (PD) or phosphorothioate (PT) backbones were assayed for their ability to inhibit the interaction of V3 loop specific Mabs with the gpl20 molecule: SEQ. ID. NOs. 31 (1173) and 32 (1174); SEQ ID. NOs. 24 (1100-07) and 39 (1100-21); or SEQ. ID. NOs. 42 (1229) and 43 (1230). To do this, immobilized gpl20 was preincubated with oligonucleotides before washing and the addition of Mab NEA 9284 (panel A) or Mab NEA 9301 (panel B). Figure 11 shows a schematic diagram of the HIV-1 genome not drawn to scale. Figure 12 shows analysis of DNA (PCR) and RNA (RT-PCR) extracted from SUP Tl cells three days post-infection with H1V-1. (Left Panel). PCR analysis of HIV-1 infected drug treated SUP Tl cell DNA used 0.1 μg of total extracted DNA for each reaction. In this experiment either AZT, at 0.3 μM which is 10 fold over the ED^ value (lane 1) or 1100-15 at 5.0 (lane 2) or 0.3 μM (lane 3) were added to SUP Tl cells at the same time as HIV-1. Lanes 4 (AZT), 5 (5.0 μM 1100-15) and 6(0.3 μM 1100-15) are the results of DNA samples obtained from cells in which drug was added 8 hours post-infection. Lanes 8 to 10 contain 10, 100 or 1000 ng of DNA extracted from HIV-1 infected control SUP Tl cells. The band corresponding to 220 bp is the predicted size of the internal β-actin control and the 200 bp fragment is the predicted size for the amplified portion of the HIV-1 genome. The right panel contains RT-PCR analysis of extracted RNA (1 μg/reaction) obtained from cells treated in an identical fashion as those described in lanes 1-6 of the left panel. Lanes 7 and 8 are control HIV-1 infected cell mRNA and lanes 9 and 10 are the results obtained using uninfected untreated SUP Tl cell mRNA. Figure 13 shows the results of three oligonucleotides (lO^M) incubated with increasing concentrations (0,7.5,15,30,60 and 120 mM) of KC1 (lanes 1-6 for 1100-15, 7-12 for 1100-18 and 13-18 for Z106-50). The nucleotide markers are poly dT. Figures 14A and 14B show a line model and space filling model for 1100-15. A line model (14A) of 1100-15 folded into an intramolecular tetrad of the Oxytricha class is depicted. The 5'-end of the molecule is in the bottom left hand side. The bases (Gs) are stacked on top of each other with the 4 bases in each plane stabilized through their hydrogen bonding with each other and their interaction with the K + ion complex in the center of the tetrad. The space filling model (14B) also has the 5' bases in the lower left hand corner. The lighter colored atoms are part of the G-tetrad and the darker shaded atoms are part of the loop structures. The K + ion is buried within the tetrad. Figure 15 displays a KC1 titration. DETAILED DESCRIPTION OF THE INVENTION It is readily apparent to one skilled in the art that various substitutions and modifications may be made to the invention disclosed herein without departing from the scope and spirit of the invention. The term "oligonucleotide" as used herein is defined as a molecule comprised of two or more deoxyribonucleotides or ribonucleotides, preferably more than ten. Its exact size will depend on many factors including the specificity and anti- viral activity of the oligonucleotide for various viruses. In addition, bases can refer to unnatural (synthetic) bases used in place of an A, C, T or G. In referring to "bases" herein, the term includes both the deoxyribonucleic acids and ribonucleic acids. The following abbreviations are used. "A" refers to adenine as well as to its deoxyribose derivative, "T" refers to thymine, "U" refers to uridine, "G" refers to guanine as well as its deoxyribose derivative, "C" refers to cytosine as well as its deoxyribose derivative. A person having ordinary skill would readily recognize that these bases may be modified or derivatized to optimize the methods of the present invention. In addition, bases can refer to unnatural (synthetic) bases used in place of an A, C, T, or G. The term "inhibition" of viral replication is meant to include partial and total inhibition of viral replication as well as decreases in the rate of viral replication. The inhibitory dose or "therapeutic dose" of the compounds in the present invention may be determined by assessing the effects of the oligonucleotide on viral replication in tissue culture or viral growth in an animal. The amount of oligonucleotide administered in a therapeutic dose is dependent upon the age, weight, kind of concurrent treatment and nature of the viral condition being treated. The term "pharmacological dose" as used herein refers to the dose of an oligonucleotide which causes a pharmacological effect when given to an animal or human. The pharmacological dose introduced into the animal or human to be treated, will provide a sufficient quantity of oligonucleotide to provide a specific effect, e.g., (1) inhibition of viral protein or enzymes, (2) inhibition of viral-specific replication, (3) preventing the target site from functioning or (4) damaging the duplex DNA at the specific site or (5) ablating the DNA at the site or (6) inhibiting the transcription/translation of the gene under the regulation of the site being bound or (7) internal inhibition of transcription or translation of the gene containing the sequence. One skilled in the art will readily recognize that the dose will be dependent upon a variety of parameters, including the age, sex, height and weight of the human or animal to be treated, the organism or gene location which is to be attacked and the location of the target sequence within the organism. Given any set of parameters, one skilled in the art will be able to readily determine the appropriate dose. The term "pathophysiological state" as used herein refers to any abnormal, undesirable or life-threatening condition caused directly or indirectly by a virus. The term "GTOs" means an oligonucleotide in which there is a high percentage of deoxyguanosine, or contains two or more segments (runs) of two or more deoxyguanosine residues per segment. As used herein, the term "guanosine tetrads" refers to the structure that is formed of eight hydrogen bonds by coordination of the four O 6 atoms of guanine with alkali cations believed to bind to the center of a quadruplex, and by strong stacking interactions. Of particular interest to the 1100-15 class of GTO is the structure of the telomere sequence repeat T 4 G 4 , first detected in Oxytricha. The oxytricha repeat has been studied in oligonucleotides by NMR and by crystallographic methods. See Smith et al., Nature 1992, 356:164-68, and Kang et al., Nature 1992 356:126-31. As predicted from numerous previous physical and biochemical studies, both the NMR and crystallographic studies suggest that folding is mediated by square planar Hoogsteen H-bonding among G-residues, with overall antiparallel orientation of the four strand equivalents comprising the tetrad fold. As expected, the crystallography has shown that the structure is selectively stabilized by tight binding of a small monovalent cation to the O 6 oxygen of guanosine. The present invention provides a method of treating a pathophysiological state caused by a virus, comprising the step of administering a pharmacological dose of an oligonucleotide, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains sufficient contiguous guanosine so that a guanosine tetrad (inter- or intra- molecular) can form, and the three dimensional structure of the oligonucleotide is stabilized by guanosine tetrads formed at strategic locations. Generally, this method of treating a virus-induced pathophysiological state may be useful against any virus. More preferably, the methods of the present invention may be useful in treating pathophysiological states caused by viruses such as herpes simplex virus, human papilloma virus, Epstein Barr virus, human immunodeficiency virus, adenovirus, respiratory syncytial virus, hepatitis B virus, human cytomegalovirus and HTLV I and II. Generally, the oligonucleotides of the present invention contain a percentage of guanosine bases high enough to ensure anti- viral efficacy. The guanosine is important in forming tetrads which stabilize the three dimensional structure of the Oligonucleotides. Thus, the oligonucleotides of the present invention may have any percentage of guanosine bases which will allow for tetrad formation provided that the oligonucleotide exhibits anti- viral activity. Preferably, the oligonucleotides of the present invention contain two or more segments of two or more guanosine bases, and an overall high percentage of G. Generally, the oligonucleotides of the present invention may be capped at either the 3' or the 5' terminus with a modifier. Preferably, the modifier is selected from the group consisting of polyamine or similar compounds that confer a net positive charge to the end of the molecule, poly-L-lysine or other similar compounds that enhance uptake of the oligonucleotide, cholesterol or similar lipophilic compounds that enhance uptake of the oligonucleotide and propanolamine or similar amine groups that enhance stability of the molecule. The phosphodiester linkage of the oligonucleotides of the present invention may be modified to improve the stability or increase the anti-viral activity. For example, the phosphodiester backbone of the oligonucleotide may be modified to a phosphorothioate linkage. Other such modifications to the oligonucleotide backbone will be obvious to those having ordinary skill in this art. The present invention also provides specific methods of treating viral states. For example, the present invention provides a method of treating a pathophysiological state caused by a herpes simplex virus, comprising the step of administering a pharmacological dose of an oligonucleotide, said dose being sufficient to inhibit the replication of said virus, wherein the three dimensional structure of said oligonucleotide is stabilized by the formation of guanosine tetrads. Also provided in the instant application is a method of treating a pathophysiological state caused by human immunodeficiency virus, comprising the step of administering a pharmacological dose of an oligonucleotide, said dose being sufficient to inhibit the replication of said virus, wherein the three dimensional structure of said oligonucleotide is stabilized by the formation of guanosine tetrads. The present invention also provides a method of treating a pathophysiological state caused by human papilloma virus, comprising the step of administering a pharmacological dose of an oligonucleotide, said dose being sufficient to inhibit the replication of said virus, wherein the three dimensional structure of said oligonucleotide is stabilized by the formation of guanosine tetrads. In addition, the present invention provides a method of treating a pathophysiological state caused by human cytomegalovirus, comprising the step of administering a pharmacological dose of an oligonucleotide, said dose being sufficient to inhibit the replication of said virus, wherein the three dimensional structure of said oligonucleotide is stabilized by the formation of guanosine tetrads. The present invention also provides a method of treating a pathophysiological state caused by hepatitis B virus, comprising the step of administering a pharmacological dose of an oligonucleotide, said dose being sufficient to inhibit the replication of said virus, wherein the three dimensional structure of said oligonucleotide is stabilized by the formation of guanosine tetrads. This invention discloses a novel anti-viral technology. The total number of antiviral mechanisms by which oligonucleotides, and especially G-rich oligonucleotides, work is not completely known. However in the different virus culture systems listed above, G-rich oligonucleotides were able to significantly reduce virus production in each, and the present invention is drawn to oligonucleotides that have three dimensional structures stabilized by the formation of guanosine tetrads. Prior to this invention, investigators tried to establish anti-viral mechanism based on anti-sense technology, and yet the present invention shows that oligonucleotide exhibition of anti-viral activity may not be via inhibition of translation events. The present invention demonstrates poly and/or oligonucleotides inhibit growth of HIV-1, HSV1, HSV2, FMLV and HCMV and other viruses if the molecule contains a high percentage of ribo- or deoxyriboguanosine. The rest of the molecule is composed of thymine, cytosine, xanthosine or adenine nucleotides (ribo- or deoxyribo-), their derivatives, or other natural or synthetic bases. The 5' and 3' termini of the oligonucleotide can have any attachment which may enhance stability, uptake into cells (and cell nuclei) or anti-viral activity. The backbone which connects the nucleotides can be the standard phosphodiester linkage or any modification of this linkage which may improve stability of the molecule or anti-viral activity of the molecule (such as a phosphorothioate linkage). Structural formulas for several different G-rich oligonucleotides disclosed in the instant invention are listed below in Table 1. All oligonucleotides listed here have a propanolamine group attached to their 3' end. This amine group confers stability against nuclease digestion to these oligonucleotides. TABLE 1. SEQ ID NO 5(B106-62) 5'-gtggtggtggtgttggtggtggtttggggggtgggg-3' SEQ ID NO 6(B106-71) 5'-gtggttggtggtggtgtgtgggtttggggtgggggg-3' SEQ ID NO 21(1100-01) 5'-tggtgggtgtgtggggggtgttgggggttgttggtggggtggtgg-3' SEQ ID NO 24(1100-07) 5'-gtggtgggtgggtgggtggtgggtggtggttgtgggtgggtggtg-3' SEQ ID NO 28(1100-50) 5'-ggtggtggggtggttgttgggggttg-3' SEQ ID NO 29(1100-51) 5'-ggtggtggggtggttgttgggggttgttgggggtgtgtgggtggt-3' SEQ ID NO 26(1100-11) δ'-gatccatgtcagtgacactgcgtagatccgatgatccagtcgatg-S' SEQ ID NO 12(G101-50) 5'-ggtgggtggtttgtgtggttggtgggtttt-3' SEQ ID NO 13(G105-50) 5'-ggggggggggtgtgggggggggttgtggtgg-3' SEQ ID NO 14(G106-50) 5'-ggtgggtgggttggggggtgggtgggg-3' SEQ ID NO 15(G109-50) 5'-tggggtttgggtggggggttgggtggttg-3' SEQ ID NO 16(G110-50) 5'-gggtggtggtgttggtgttgtgtg-3' SEQ ID NO 17(G113-50) 5'-ggtgggggggttggtgtgtttg-3' SEQ ID NO KA100-00) 5'-tgggtggggtggggtgggggggtgtggggtgtggggtg'3' SEQ ID NO 2(A100-50) 3'-tgggtggggtggggtgggggggtgtggggtgtggggtg-5' SEQ ID NO 4(A101-00) 5'-ggtggtgggggggggtggggtggtggtgggggtgttgg-3' SEQ ID NO 18(HIV26ap) 5'-gtgtgggggggtggggtggggtgggt-3' SEQ ID NO 19(HIV26ctl) 5'-gggtgggtgggtgggtgggtgggtgg-3' SEQ ID NO 9(B107-51) 5'-ggtggggtggtggtggttggggggggggggt-3' SEQ ID NO 10(B133-55) 5'-ggtggttggggggtggggggg-3' SEQ ID NO 1KB133-55) 5'-gggtggggtggtgggtggggg-3' SEQ ID NO 20(1100-00) 5'-gttgggggttgttggtggggtggtgg-3' SEQ ID NO 27(1100-12, PT) 5'-gttgggggttgttggtggggtggtgg-3' SEQ ID NO 22(1100-05) 5'-tggtgggtgtgtggggggtgttgggggttgttggtggggtggtgg- CHOL SEQ ID NO 23(1100-06) 5'-gtggtgggtgggtgggtggtgggtggtggttgtgggtgggtggtg- CHOL SEQ ID NO 25(1100-08) 5'-gttgggggttgttggtggggtggtgg-CHOL SEQ ID NO 3 5'-gggtgggtgggtgggtgg-3' SEQ ID NO 30 5'-gggtggttgggtggttgg-3' SEQ ID NO 31(1173) 5'-gggtgggtgggtgggtgg-3' SEQ ID NO 32(1174, PT) 5'-gggtgggtgggtgggtgg-3' SEQ ID NO 33(1100-15) 5'-gtggtgggtgggtgggt-3' SEQ ID NO 34(1100-16) 5'-gtggtgggtgggtgggtggtgggtggt-3' SEQ ID NO 35(1100-17) 5'-gtggtgggtgggtgggtggtgggtggtggttgtgggt-3' SEQ ID NO 36(1100-18) 5'-ttgtgggtgggtggtg-3' SEQ ID NO 37(1100-19) 5'-tggtgggtggtggttgtgggtgggtggtg-3' SEQ ID NO 38(1100-20) 5'-gtgggtgggtggtgggtggtggttgtgggtgggtggtg-3' SEQ ID NO 39(1100-21, PT) 5'-gtggtgggtgggtgggtggtgggtggtggttgtgggtgggtggtg-3' SEQ ID NO 40(1231) 5'-gatccatgtcagtgacac-3' SEQ ID NO 41(1232, PT) 5'-gatccatgtcagtgacac-3' SEQ ID NO 42(1229) 5'-cccccccccccccccccc-3' SEQ ID NO 43(1230, PT) 5'-cccccccccccccccccc-3' SEQ ID NO 44(1198) 5'-ttcatttgggaaacccttggaacctgactgactggccgtcgttttac-3' SEQ ID NO 45(1200) 5'-gtaaaacgacggcca-3' SEQ ID NO 46(1100-25) 5'-gtggtgggtgggtgggg-3' SEQ ID NO 47(1100-26) 5'-gtggtgggtgggtggg-3' SEQ ID NO 48(1100-35) 5'-tggtgggtgggtgggt-3' SEQ ID NO 49(1100-27) 5'-gtggtgggtgggt-3' SEQ ID NO 50(1100-28) 5'-gtggtgggt-3' SEQ ID NO 51(1100-30) 5'-gtgggtgggtgggt-3' SEQ ID NO 52(1100-29) 5'-gtgggtgggt-3' The present invention also provides novel guanosine-rich oligonucleotides having anti-viral activity. The following examples are offered by way of illustration and are not intended to limit the invention in any manner. EXAMPLE 1 HSV-2 CULTURE ASSAY In viral yield reduction assays, Vero cells (4 x 10 4 cells/tissue culture well) were incubated with oligonucleotide (s) for 14 hours before the oligonucleotide was removed and virus (HSV-2 strain HG52) added to the cells at a multiplicity of infection (m.o.i.) of 0.1 to 1.0 (4 x 10 3 to 4 x 10 4 PFU). The infection was allowed to proceed for 10 minutes after which the cells are washed and fresh media, containing the same oligonucleotide is added for an additional 14 hours. Then the cells are subjected to a freeze/thaw lysis after which the released virus is titered. EXAMPLE 2 HIV-1 CULTURE ASSAY The SUP Tl T lymphoma cell line was infected with HIV-1 strain DV at a multiplicity of infection (m.o.i.) of 0.1 for one hour at 37°C. After the infection, free virus was washed off and the newly infected cells were plated (5 x 10 4 cells) in quadruplicate in 96 well plates that had been prepared with various dilutions of oligonucleotide. The final concentration of drug varied between 0.1 and 20 uM. After 3 days of incubation at 37°C, the plates were scored for the presence of multinucleated giant cells (syncytia). In assays designed to inhibit syncytia formation, a number of oligonucleotides exhibited anti-HIV-1 activity. The oligonucleotides and their ED50 are listed in Table 2. 1100-05 is the same as 1100-01 with a cholesterol group attached to the 3' end via a triglycyl-linker. 1100-08 is the same as 1100-00 with a cholesterol group attached to the 3' end via a triglycyl-linker. 1100-07 was designed as a sequence isomer to 1100-01 and 1100-06 is the cholesterol derivative of 1100-07. AlOO-00 is the same sequence in the opposite orientation to HIB38p (A100-50). 1100-07, originally designed as a control for 1100-01 to be used in anti-FMLV experiments, was the most efficacious oligonucleotide tested against HIV-1. In other experiments, the HIV-1 strain LAV was used to infect MT-2 cells at an m.o.i of 0.01. After 7 days, these cells were scored for cytopathic effects (CPE). In anti-HIV-1 assays in which MT-2 cells were infected at an m.o.i. of 0.01, several G-Rich oligonucleotides were able to inhibit viral- induced cytopathic effects with effective dose 50's (ED50s) in the 0.5-1.0 uM range (Figure 3). The oligonucleotides shown in Figure 3 were effective in the 0.5 to 1.0 uM range, including AlOO-00 (HIV38p) and A100-50 (H_V38ap), AlOl- 00 (H_V38ctl), HIV-26ctl. The oligonucleotide HIV-26ap exhibited less efficacy in this assay with an ED50 in the 5 to 10 uM range. In Figure 3, TE represents buffer alone, i.e., no drug, while AZT and ddC are control drugs. TABLE 2 EDg n for oUgonucleotides in an anti-HIV-1 syncytia formation assay. G-Rich oligonucleotide ED50 1100-00 3.75 μM. 1100-01 4.50 μM 1100-05 3.25 μM 1100-08 3.25 μM 1100-06 0.70 μM 1100-07 0.25 μM AlOO-00 3.25 μM EXAMPLE 3 FMLV CULTURE ASSAY Friend Murine Leukemia Virus (FMLV) was grown in a chronically infected murine fibroblast cell line (pLRB215) or can be propagated in an acute assay system by infection of NIH3T3 cells. When the chronically infected cell line was used, pLRB215 cells were split (1 x 10 5 ) into 24 well culture dishes and incubated 16 to 20 hours at 37°C. The media was then removed and replaced with media containing various concentrations of oligonucleotide. After 1, 3 or 5 days, culture media was assayed for the presence of the viral reverse transcriptase enzyme. In acute assays, NIH3T3 cells were split (1 x 10 4 ) into 96 well dishes and allowed to incubate for 16-20 hours. After incubation, culture media was removed and concentrated virus stock (10 ul) was added to each well in 100 ul of completed media containing 2 ug/ml polybrene. The virus infection was allowed to proceed for 18 hours at which time the virus containing media was removed and complete media containing various concentrations of oligonucleotide was added. After 4 to 7 days, the culture media was assayed for the presence of viral reverse transcriptase. EXAMPLE 4 HCMV CULTURE ASSAY Human cytomegalovirus was cultured in the human diploid lung fibroblast cell line MRC-5. These cells were split and placed into 24 well culture dishes and preincubated for 24 hours with various concentrations of oligonucleotide (0.5 to 20 uM) in complete media. The oligonucleotide was then washed off and virus was added to the cells (approximately 0.1 m.o.i.) for 2 hours at 37 C C. The virus was then removed and complete media containing the same concentration of oligonucleotide was added. Cells were then placed at 37°C for 10-12 days at which time virus in the culture media was titered using a standard agar overlay procedure. EXAMPLE 5 BACTERIAL T3 AND T7 ASSAYS In this assay system, a 2 kb fragment (Hindlll to Ecor Rl) of the FMLV virus (clone 57) was molecularly cloned between the Hind III/Eco Rl sites 10 bp downstream of the bacterial T7 promoter (p275A) or 50 bp downstream of the bacterial T3 promoter (pBSFMLV2). A schematic representation of these two recombinant plasmids can be seen in Figure 1. Isolated recombinant DNA was then digested with Dde I. Oligonucleotides were incubated then with the digested DNA and then subjected to in-vitro transcription using either the T7 or T3 bacterial enzymes. EXAMPLE6 REVERSETRANSCRIPTASEASSAY In this assay, reverse transcriptase (either MMLV which is commercially available or FMLV from pLRB215 culture media) was incubated with various concentrations of oligonucleotide and then assayed using the enzyme linked oligonucleotide sorbent assay (ELOSA), the ELOSA kit is commercially available from New England Nuclear. EXAMPLE 7 EUKARYOTIC IN VITRO TRANSCRIPTION In this assay, a recombinant plasmid containing the HSV-1 IE 175 promoter fused to the bacterial chloramphenicol acetyltransferase gene (CAT) was linearized and used as a template for run off transcription studies. The commercially available HeLa cell nuclear extracts or prepared nuclear extracts of HSV-2 infected VERO cell were used. EXAMPLE 8 INHIBITION OF HSV-2 ACTIVITY The oligonucleotide B106-62 was originally designed to form a triple helix structure with a portion of the promoter region of the major immediate early protein of HSV-2 (IE 175). The phosphorothioate derivatives of these two oligonucleotides were synthesized and tested for anti- viral activity against HSV-2. Figure 2 shows that the B 106-62 oligonucleotide at 20 μM was able to reduce viral titers by approximately 20% whereas the phosphorothioate version (B 106-96) reduced virus by 50% in the submicromolar concentration range. The control oligonucleotide (B 106-97), the phosphorothioate backbone derivative of B106-71, was also able to inhibit virus at the same levels as B106-96. Even when an extensive washing procedure at a pH of 3 was employed to remove excess virus not internalized during the infection, incubation with both B106-96 and B106-97 were able to significantly reduce virus yield. Thus, the mechanism of anti- viral activity was not just blocking adsorption of HSV-2 virions to cells. Figure 2 also shows the results of acyclovir in the same molar range as the oligonucleotides. Acyclovir was tested against two different stocks of HSV-2 strain HG52, as illustrated in Figures 4a and 4b. EXAMPLE 9 A. OUGONUCLEOTIDE SYNTHESIS The 47-mer DNA template oligonucleotide used was SEQ.ID. NO. 44 (1198) and the 15-mer primer sequence was SEQ.ID. NO. 45 (1200). All oligonucleotides used in these examples were synthesized on a DNA synthesizer (Applied Biosystems, Inc., model 380B or 394), using standard phosphoramidite methods. All oligonucleotides were synthesized with an amino modified 3 '-terminal, which results in the covalent attachment of a propanolamine group to the 3'-hydroxyl group or with a cholesterol moiety attached to the 3'-terminal via a triglycyl-linker. Oligonucleotides used in this example were capped at their 3 '-terminal with either a propanolamine or a cholesterol moiety to reduce degradation by cellular exonucleases. Phosphorothioate containing oligonucleotides were prepared using the sulfurizing agent TETD. The 3'-cholesteryl modified oligonucleotides were prepared and purified as described by Vu et al. (in Second International Symposium on Nucleic Acids Chemistry, Sapporo, Japan, 1993). B. STABILITY AND TOXICITY Oligonucleotides with either full length phosphodiester (PD) or full length phosphorothioate (PT) backbones were stable in the culture media for 4 days while oligonucleotides consisting of a more random composition of nucleotides were rapidly degraded. This indicates that the 3'-modified G-rich oligonucleotides with PD backbones were stable against both endonuclease and exonuclease digestion over a defined four day incubation in culture. The concentration of oligonucleotide needed to reduce cell proliferation by 50% (TCg o ) of selected compounds, based on the dye metabolism assay was approximately 40 to 50 μM for oligonucleotides with PD backbones and 15 to 40 μM for those compounds containing a PT backbone. The TC^ for selected oligonucleotides are presented in Table 3. TABLE 3 βuanosine/thymidine and control oligonucleotide sequences Oligo" Length 3' -Modification 11 Sequence TC _,___ 5' -gtggtgggtgggtgggtggtgggtggtggttgtgggtgggtggtg-3' >50 μM 5' -gtggtgggtgggtgggtggtgggtggtggttgtgggtgggtggtg-3' 5' - gttgggggttgttggtggggtggtgg-3' 37 μM 5' - gttgggggttgttggtggggtggtgg-3' 5' - gttgggggttgttggtggggtggtgg-3' 18 μM 5' -tggtgggtgtgtggggggtgttgggggttgttggtggggtggtgg-3' 5' -tggtgggtgtgtggggggtgttgggggttgttggtggggtggtgg-3' 5' -tgggtggggtggggtgggggggtgtggggtgtggggtg -3' 5' -gggtgggtgggtgggtgg -3' 5' -gatccatgtcagtgacactgcgtagatccgatgatccagtcgatg-3' 46.5 μM 5' -gatccatgtcagtgacac -3' 5' -cccccccccccccccccc -3' All oligonucleotides listed were synthesized with phosphodiester backbones except 1100-12 which had phosphorothioate (PT) linkages. The capping group at the 3' -end of the oligonucleotide was either a propanolamine or cholesterol moiety. Median inhibitory (toxic) concentration in tissue culture. EXAMPLE 10 INHIBITION OF HIV-1 PRODUCTION IN AN ACUTE ASSAY SYSTEM: A. Cytotoxicity and Stability Assays. The cytotoxicity of selected oligonucleotides was assayed using the CellTiter 96™ Aqueous Non-Radioactivity Cell Proliferation Assay (Promega) . This is a colormetric method for determining the number of viable cells in proliferation or chemosensitive assays using a solution if MTS. Dehydrogenase enzymes found in metabolically active cells convert MTS into a formazan product. The SUP Tl cells used in the cytotoxicity assays were in log phase growth at the time of the assay. Cytotoxicity profiles for GTOs with PD backbones such as 1100-15 had TC^s (50% cytotoxic concentration) in the range of 30 to 50 μM while GTOs with PT backbones such as 1100-15 had TC so S in the 10 to 30 μM range. The TO* for AZT in this assay format was approximately 10 μM . Blockage of the hydroxyl terminus of oligonucleotides has been shown by many investigators to greatly reduce degradation by cellular exonucleases therefore all oligonucleotides used in these studies were modified at their 3'- end with either a propanolamine group or a cholesterol group. For stability studies, 10 μM of GTOs were incubated in MEM (GIBCO) supplemented with 10% FBS. Aliquots were taken after 10 min, 1 day, 2 days, 3 days and 4 days. The aliquots at each time point were immediately extracted twice with 50:50 phenol-chloroform solution and then precipitated by the addition of ethanol. The recovered oligonucleotides were 5'-end-labeled using [γ- 32 P]ATP and polynucleotide kinase. The integrity of the oligonucleotides was then analyzed on a 20% polyacrylamide gel with 7 M urea. The results indicate that a portion of each GTO with a PD backbone was present in the culture medium of three to four days while oligonucleotides composed of a more random assortment of all four nucleotides were rapidly degraded. In addition, positions within PD GTOs where there exist two or more contiguous pyrimidines are more susceptible to endonuclease digestion than regions containing purines or alternating purines and pyrimidines. B. Long term suppression of Acute HIV-1 infections in SUP Tl cells. The anti-HIV-1 activity of a series of GTOs, with PD backbones, containing different sequences motifs was tested. One of the sequence motifs tested (oligonucleotide 1100-07) was 10 fold more active at inhibiting HIV-1 induced syncytium formation than the other motifs tested (e.g. 1100-00 shown in Table 1). 1100-07 and its derivatives (length and chemical modification) were further tested for their ability to inhibit virus in a dose-dependent fashion by measurement of syncytium formation and viral p24 production. Briefly, H_V-1 DV was used to infect the SUP Tl lymphoblastoid cell line at a multiplicity of infection (m.o.i.) of 0.1 TCIDg o for one hour at 37° C prior to washing and resuspension in increasing concentrations of GTOs. The cells (2 x 10 4 cells/well) were inoculated in triplicate in 200 ul of RPMI 1640 containing 10% fetal calf serum. Four days later, the number of syncytia per well or the level of p24 in the medium was determined. The results of these assays are presented in Table 4. TABLE 4. Ouanosine/thymidine oligonucleotide sequences . Oligo Length linkage" Sequence 1100 -07 45 mer PD 5'- gtggtgggtgggtgggtggtgggtggtggtlgtggglgggtggtg -21 45 mer PT 5'- gt-^gext tgggt gt tggt ttgtgggtg gtggtg -20 38 mer PD 5'- gtggglggglggtgggtgglggltgtgggtgggtggtg -19 29 mer PD - ■ '- tggtgggt tggttgt tgg tggtg -18 16 mer PD 5'- ttgtgggtgggtggtg -17 37 mer PD 5'- gtggtg_^g_a_tgggtggtgggtggtggttgtgg8t -16 27 mer PD 5^-gtggtgggtgggtgggtggtgggtggt I -15 17 mer PD 5'-gtggtgggtgggtgggt to _υ -00 26 mer PD 5'-gttgggggttgttggtggggtggtgg -12 26 mer PT 5'-gttgggggttgttggtggggtggtgg AZT 0.04 0.40 >200 a The internucleotide backbone linkages are denoted as PD for phosphodiester and PT for phosphorothioate. b The ED50 values for the syncytium and p24 inhibition assays in uM concentrations. c T.I. = therapeutic index. The experimental results indicated that GTOs with simple PD linkages were capable of inhibiting HIV-1 syncytia formation and p24 production in culture. To determine the effect of backbone modification on GTO anti-viral activity, the PD backbone in two oligonucleotides sequences motifs was replaced with a PT backbone. The phosphorothioate containing oligonucleotides (1100-12 and 1100-21) where then tested for their ability to inhibit HIV-1 induced syncytium formation and production of HIV-1 p24 in the SUP Tl acute assay system (Table 4). The results from these assays indicated that the presence of sulfur molecules in the oligonucleotide backbone greatly enhanced the anti-viral activity of 1100-00 (1100-12) but had little if any effect on 1100-07 (1100-21) (Table 4). It was apparent from our studies that the anti-viral activity of 1100-07 was maintained when we reduced the length of the molecule to 17 by deleting segments from the 3'-end (1100-15, -16, -17) but not by deletions from the 5'- end (1100-18, -19, -20). To further determine to optimal size of the PD oligonucleotide needed for maximal anti-HIV-1 the 1100-15 size variants listed in Table 5 were synthesized and assayed for antiviral activity. TABLE 5. Inhibition of HIV-1 Induced Syncytia Using Size variants of 1100-15. AZT 0.02 * At 5 uM these compounds suppressed virus at least 7 days post-removal of drug. All other compounds at 5 uM were the same as AZT 7 days after removal of drug. The duration of the viral suppression was assayed by changing the medium in HIV-1 infected cultures containing 2.5 uM of various oligonucleotides to complete media without added oligonucleotide on day 4 post- viral infection. The production of viral p24 antigen was then assayed on day 7 and day 11 post-infection. The results of this experiment indicated that the shorter variants of 1100-07 (1100-15 and 1100-16) as well as the PT version of this molecule (1100-21), were capable of totally suppressing H1V-1 p24 production for at least 7 days after removal of the drug from the culture medium (Table 6). This substantial level of prolonged inhibition was >99% for 1100-15, 1100-16 and 1100-21 when compared to the p24 antigen levels obtained for untreated HIV-1 infected cells (Table 6). The quantitation of p24 production relative to untreated HIV-1 infected SUP Tl cells for all oligonucleotides tested is presented in Table 6. The presence of sulfur molecules in the backbone of oligonucleotide 1100-07 (1100-21) had a more marked effect on the reduction of virus seven days after removal of compound from the culture medium than was observed at the four day post-infection assay point (Table 5). TABLE 6. Detection of HIV-1 p24 antigen in the culture media of GTO treated SUP Tl cells. Oligonucleotide (2.5uM) Control SUP Tl cells 1100-07 1100-21 (PT) d 1100-15 1100-16 1100-18 1100-19 1100-12 (PT) a Level of detectable p24 in culture medium relative to control (infected but untreated SUP Tl cells after subtraction of background values. b Day 4 post-infection culture medium was replaced with fresh medium without oligonucleotide. c SUP Tl cells infected with HIV-1 but not treated with oligonucleotides or AZT were used as positive control cells in this experiment. d 1100-21 and 1100-12 contain phosphorothioate backbone linkages (PT). In control experiments the culture medium from HIV-1 infected SUP Tl cells treated with AZT (4 μM) was also replaced on day 4 post-infection with drug free media. In these experiments, two days after removal of AZT from the culture medium the presence of syncytium was observed in the HIV- 1 infected cell cultures and by day 4 all cells were visibly infected with HIV-1. To determine whether the prolonged suppression of HIV- 1 was due to toxicity of the oligonucleotides, SUP Tl cells were counted for all treated samples 7 days after removal of the oligonucleotides from the infected cell cultures. The results indicated that for cells treated with 2.5 μM of drug there was no difference in the number of cells when compared with control cultures (uninfected, untreated) of SUP Tl cells. C. Inhibition of HIV expression in patient derived peripheral blood mononuclear cells (PBMCs). 1100-15 was assessed for activity in PBMC cultures derived from AIDS patients. Briefly, PHA activated uninfected PBMCs were added to 4PBMC's derived from patients with HIV infection in the presence of varying concentrations of oligonucleotide. Anti-HIV activity was assessed by analyzing supernatants, collected every three days from these mixed cultures, for the presence of HIV p24. The PHA activated PBMCs were grown in the presence of 10 units/ml of IL-1 and medium was exchanged every three days for a period of three weeks. HIV p24 antigen production was assayed in drug-treated as compared to untreated control specimens. It is interesting to note that the results in these experiments (Figure 9) observed for AZT were obtained when AZT was used at 12 uM which is roughly 300 fold greater than the EDg. for this compound. D. In-Vitro inhibition of HIV-1 reverse transcriptase (RT). The ability of oligonucleotides to inhibit HIV-1 RT in vitro has been well documented. Marshall et al. have described a competitive interaction at the active site as the mechanism by which mono- or diphosphorothioate containing oligonucleotides inhibit HIV-1 RT independent of whether the molecule tested was antisense, a random sequence or poly SdC. To determine whether 1100-15 or its parent molecule, 1100-07 (or the PT version 1100-21), was interacting with H1V-1 RT the activity of this enzyme was assayed in the presence of various concentrations of oligonucleotides. A kinetic analysis of the resultant enzyme inhibition was conducted to determine the mechanism of inhibition. The GTOs appeared to be inhibiting the RNA dependent DNA polymerase activity of the RT enzyme by competitive inhibition at the active site of the enzyme. The K; value for all of the oligonucleotides tested is presented in Table 7. The data indicate that for all oligonucleotides tested the presence of the sulfur group in the backbone greatly enhanced the interaction between the oligonucleotides and the enzymes. The median inhibitory dose (ID s -) for these oligonucleotides were also calculated (Table 7). The ID s ,, results are based on the ability of these compounds to inhibit 10 nM of HIV RT. Short oligonucleotides (18 mers) with PD or PT backbones were assayed to determine whether the nature of the nucleotide sequence contributed to inhibition of HIV-1 RT in this assay system. Comparison of the effects of the PD versions of a GTO (1173 or 1100-15), poly dC (1229) or a random nucleotide sequence (1231) suggested that at this length none of the sequence motifs inhibited RT (Table 7). Other 18 mer PD GTO sequence motifs tested yielded similar results. Enzyme inhibition monitored by both K_ and IDg o was observed for the PT versions of these same 18 mer oligonucleotides (Table 7). The degree of enhancement of observed enzyme inhibition for all oligonucleotides tested when the sulfur group was present in the backbone, was between one to three orders of magnitude (Table 7). TABLE 7. In Vitro Inhibition of HIV-1 RT by PD and PT Oligonucleotides. OHgonucleotides Length Linkage b Ki (μM) ID50 (μM) poly dC while the 1231 and 1232 oHgonucleotides were a random sequence of all four bases (GATC). b The backbone modifications are denoted as PD for phosphodiester and PT for phosphorothioate. The results from this set of experiments demonstrate that 1100-15 is minimally inhibitory to the RNA dependent DNA polymerase activity of HIV-1 RT. The data also indicates that chemically modifying GTOs, poly dC or a random sequence oligonucleotide greatly enhances the in vitro inhibitory activity of the molecule. Therefore, chemically modified oligonucleotides such as the antiviral G-rich molecule describe by Wyatt et al. [112] has by nature a different set of characteristics from oligonucleotides with natural PD backbones. E. Inhibition of the interaction of HIV-1 gpl20 with cellular CD4. The outer envelope glycoprotein gpl20 of HIV-1 mediates viral attachment to the cell surface glycoprotein CD4 in the initial phase of HIV-1 infection. The effects of both PD and PT modified oligonucleotides on this interaction were examined using a gpl20 capture ELISA kit. The concentration of the gpl20 used in these studies (125 ng/ml) was determined to be within the linear range of the detection assay. The ability of oligonucleotides to inhibit gpl20/CD4 interactions by binding to gpl20 was determined by preincubation of the test compounds with soluble gpl20 before addition to the immobilized CD4. The results of this experiment (Table 8) are presented as the concentration of oligonucleotide needed to reduce by 50% CD4 bound gpl20 (IDg o [gpl20]). The reciprocal experiment was then performed to measure the ability of the oligonucleotides to inhibit these interactions by binding to immobilized CD4. In this set of experiments 1100-00, 1100-07 and the PT versions of these two oligonucleotides were capable of preventing the interaction of gpl20 with immobilized CD4 (IDg o [CD4], Table 8). For both sequences tested, the PT version of the oligonucleotide the IDg o values were 50 to 100 fold lower than with the PD version. A fixed length (18 mer) set of oligonucleotides with either PD or PT backbones were assayed to determine whether the nature of the nucleotide sequence contributed to inhibition of gpl20/CD4 interactions. As was observed for the inhibition of HIV-1 RT, the PD versions of these molecules had little to no measurable effects on the binding of gpl20 with CD4. However, the PT versions of these oligonucleotides did yield measurable inhibitory activity. The 18 mer GTO (1174) interrupted gpl20/CD4 interactions at approximately 10 fold lower concentrations than poly (SdC) 18 (1230) while the random sequence 18 mer (1232) had no measurable activity (Table 7). TABLE 8. In Vitro Inhibition of HIV-1 gpl20 Interaction with CD4 by PD and PT Oligonucleotides. OHgonucleotide Linkage * ID50 [gpl20](μM) ID50[CD4](μM) I 100-00 PD 3.50 18 I 100-12 PT 0.08 0.475 I 100-07 PD 0.80 4.25 1 100-21 PT 0.07 0.048 1173 PD >100 >100 1174 PT 0.09 0.45 1229 (poly dC) PD >100 >100 1230 (poly dC) PT 1.00 3.25 1231 (GATC) PD >100 >50 1232 (GATC) PT >10 >10 * Each pair of oHgonucleotides contain the same sequence and differ onlyTn the nature of their backbone linkage. b The backbone modifications are denoted as PD for phosphodiester and PT for phosphorothioate. F. Oligonucleotide interactions with the v3 loop of fflV-l gp__20. It has been reported previously that poly SdC oHgonucleotides were able to bind to the third variable loop domain of HIV-1 gpl20 (v3 loop). The degree of interaction was reported to be dependent on the length of the oligonucleotide studied, with a rapid decrease in binding affinity observed for compounds shorter than 18 nucleotides. The detection antibody used to monitor inhibition of gpl20/CD4 interactions in the capture gpl20 ELISA KIT (HRP-α-GP120) used in this study (Table 8) recognized an epitope in the gpl20 v3 loop (manufacturer's information). For this reason control experiments were performed to determine whether the observed inhibition of gpl20/CD4 interactions was due in part, or in whole, to interference with the HRP-α-gpl20 detection antibody. The results indicated that 1100-07 and 1173 (PD backbones) did not inhibit the detection of immobilized gpl20 however, the PT oligonucleotides tested (1100-21 and 1174) were able to slightly inhibit the detection of gpl20 at oligonucleotide concentrations above 5 μM. This level of inhibition was too small to account for the IDg o [gpl20] values presented for 1100-21 and 1174 in Table 8. Further analysis of oligonucleotide interactions with the v3 loop was conducted using a v3 loop specific murine Mab, NEA-9284 (figure 10). PT oligonucleotides were able to inhibit binding of NEA-9284 to gpl20. The presence of bound gpl20 specific Mab was determined using a HRP-labeled goat-α-mouse antibody. The results of these experiments indicate that PT oligonucleotides were able to inhibit binding of NEA -9284 to gpl20. The ID^ for the most active oligonucleotide (1100-21) was approximately 4 to 7 μM. This concentration is approximately 10 to 30 fold higher than the EDg o for this oligonucleotide against HIV-1 in culture (Table 8). The PD oligonucleotides tested did not inhibit the binding of any Mab to gpl20 therefore, it is unlikely that this is the mechanism by which the PD GTOs such as 1100-07 (and hence 1100-15) are inhibiting H1V-1. G. Analysis ofHIV-1 RNA and DNA in single cycle assays. Total RNA and DNA were extracted from SUP Tl cells 36 hours after infection with 0.1 m.o.i of HrV-l DV . In this assay the infected cells were treated with 1100-15 or AZT at various time points before, during or after infection. Harvesting of the infected cells at 36 hr post-infection allowed for the analysis of approximately one round of viral replication. A schematic diagram of the positions of the PCR primers used in the DNA and RNA analysis is shown in figure 11. Total extracted DNA was analyzed using a PCR primer set which would amplify a 200 bp portion of the viral genome spanning the repeat element (R) into the gag gene. The primer set detected full-length or nearly completely synthesized viral DNA. This is the last region of the minus strand of viral DNA that is synthesized. Thus, for DNA to be detected by this primer set, two template-switching events have occurred and contiguous 5'LTR to gag sequences must be present on either the minus or plus strand of DNA. In the same reaction mixture a PCR primer set which would amplify a 220 bp region of the human b-actin gene was used. The results indicate that in cells treated with AZT there was a marked decrease in viral DNA synthesis when the drug was added up to 4 hrs post-infection as expected (data in Figure 12 shows zero hour and 8 hour time of addition studies). The effects of 1100-15 on the early rounds of viral DNA synthesis was minimal. The results of this experiment indicate that 1100-15 did not inhibit virus entry into the cells because of the detectable levels of viral DNA even in samples treated with 1100-15 at the same time as virus infection (zero hour addition). Furthermore, it suggests that 1100-15 has a different mechanism of action compared to AZT. Additional experiments using alternative PCR primers, suggest that there may be alterations in the viral DNA synthesis caused by 1100-15. The observed amplification products, when primers clustered in the U3 region of the virus were used, yielded a banding pattern which was not predicted and obviously different from the infected cell control (untreated) and the AZT treated infected cell samples. RNA extracted from HIV-1 infected cells was analyzed by RT-PCR. In this assay the antisense primer of the PCR primer pairs is used with MMLV RT and extracted mRNA to synthesize cDNA strand. This cDNA is then used as templates in PCR reactions. Two RNA primer sets were used to analyze unspliced (primers rl and r2) and spliced (primers rl and r3) HIV-1 transcripts. Predicted sizes of the amplified products were 101 bp and 214 bp for the unspliced and spliced species respectively. The same β -action primers used for the analysis of the DNA samples were used as controls in this experiment. The results obtained using primer pair rl and r3 are depicted in Figure 12 above. The results of this experiment clearly indicate that a reduced level of HIV-1 specific transcript was observed in samples treated with 1100-15 in the samples treated with drug at the same time as virus infection (zero hours). It is also clear that while samples treated with AZT had reduced levels of viral cDNA, viral mRNA was still being produced. The same decrease in HIV-1 specific transcript was observed in viral infected cells treated with 1100-15 when the rl and r2 primer pair was used (data not shown). H. Structural analysis of 1100-15 and 1100-26. 1100-07, and its derivative products including 1100-15 and 1100-26, are composed entirely of deoxyguanosine (G) and deoxythymidine (T). These G-rich oligonucleotides were purified using anion exchange reverse phase HPLC. Using this procedure the oligonucleotide is purified in the presence of sodium ions. Monovalent cations are known to encourage self-associated structures for G- rich molecules, all of which involve formation of G-tetrads. The G-tetrad formation involves the formation of eight hydrogen bonds by coordination of the four O 6 atoms of guanine with alkali cations believed to bind to the center of a quadruplex, and by strong stacking interactions. The oligonucleotides purified using anion exchange chromatography then have an opportunity to form inter-or intramolecular tetrads. The tetrad structure can be strengthened by replacing the sodium ion with potassium. I. Nondenaturing gel analysis. 1100-15 (17 mer, Table 5) was analyzed using nondenaturing polyacrylamide gel electrophoresis. In this experiment, trace concentrations of radiolabeled oligonucleotide (10 "7 M) was incubated with increasing concentrations of cold oligonucleotide (up to 10^M) before gel analysis in the presence of monovalent cation. Under the gel conditions used 1100-15 migrated as a unique band faster than a random coiled (denatured) 17 mer oligonucleotide would as and did so in a concentration independent fashion (data not shown). This is in contrast to 1100-18 (16 mer, Table 52 10 fold less active than 1100-15) which appeared to migrate as multiple species in a concentration dependent fashion under the same gel conditions (data not shown). The same phenomena was observed when 10 "8 oligonucleotide (total cold and radiolabeled oligonucleotide) was incubated with increasing concentration of KC1 (Figure 13). 1100-15 migrated as a unique species at all concentrations of KC1 while 1100-18 and Z106-50 (ggttgggggttggg) migrated as multiple species. The results from these assay suggests that 1100-15 folds into an intramolecular structure while other G-rich oligonucleotides such as 1100-18 and Z106-50 aggregate into higher order intermolecular structures. It is interesting to note that the PT G-rich compound described by Wyatt et al., P.N.A.S. 1994 91:1356-66, with the sequence T 2 G 4 T 2 , was claimed to fold into an intermolecular tetrad. Therefore, 1100-15 (PD backbone) is structurally and chemically different from ISIS PT oligonucleotide. J. Tetrad Structure. Principally due to its role in telomere formation, the structure of four stranded nucleic acid tetrads has been well studied. Most eukaryotes possess a repeating G-rich sequence of the form (T/C)nGm where n= 1-4 and m= 1-8. Of particular interest to the study of the 1100-15 class of GTO is the structure of the telomere sequence repeat T 2 G 4 , first detected in Oxytricha. The oxytricha repeat has been studied in oligonucleotides by NRM, Smith et al., Nature et al, Nature 1992, 356:164-68, and by crystallographic methods, Kang et al. Nature, 1992 356:126-31. As had been predicted from numerous previous physical and biochemical studies, both the NMR and crystallographic studies suggest that folding is mediated by square planar Hoogsteen H-bonding among G residues, with overall antiparallel orientation of the four strand equivalents comprising the tetrad fold. As expected, the crystallography has shown that the structure is selectively stabilized by tight binding of a small monovalent cation to the O 6 oxygen of guanosine. But surprisingly, both NMR and crystalograpy confirm that the folded structure possess alternating syn/anti glycosidic bond angles (as opposed to all anti for most duplex structures). Feigon and colleagues have used NMR and modelling to deduce the structure of a 28 base-long oligonucleotide (G 4 T 4 G 4 T 4 G 4 T 4 G 4 ,Oxy 3.5) which is capable of forming a well-defined all-antiparallel intramolecular tetrad, Smith et al., Nature 1992 356:164-68. If the GTO 1100-15 were to fold to form a stable intramolecular tetrad its NMR properties would be expected to be similar to those of the Oxy 3.5 molecule. In the folded state, the salient NMR characteristics of the intramolecular Oxy 3.5 tetrad were as follows: 1. Narrow linewidths, indicative of monomer formation only. 2. Induction of well-defined guanosine Nl Hoogsteen imino resonances in the 11.2 to 11.7 ppm range. The chemical exchange rate of these protons is very slow, reflective of the high positive cooperativity of tetrad folding and dissociation. 3. Spectral simplicity, indicative of a single predominant folded structure, rather than an equilibrium among different folded structures. 4. Intrabase H8-C1' and interbase H7-C2" NOE connectivity which demands a pattern of alternating syn-anti glycosidic bond angle throughout the "tetrad stem" of the folded structure. K. One dimensional NMR analysis. Displayed in figure 15 is a line model for 1100-15, folded to form an intramolecular tetrad of the Oxytricha class. From a physical perspective, the possibility that an intramolecular tetrad structure might form in high KC1 or NaCl is not surprising. What is surprising is the fact that this model proposes a stem region comprising a single G-octet and intervening loop regions which are only two bases long. In order to test the general feasibility of this model, a detailed 3D molecular model for a 1100-15 has been constructed. In this we have assumed that the 8 G's comprising the octet core of the structure form a standard square planar octet,and that glycosidic angles are as in the crystal and NMR structures of the antiparallel Oxytricha tetrads, Smith et al., Nature 1992 356:164-68, and Kang et al., Nature 1992 356:126-31. Additionally, a single K + ion has been introduced into the center of the G-octet, with octahedral coordination to G0 6 . Initially, 2 base loop structures were created so as to connect elements of the octet without disruption. Subsequent to this initial guess, the structure was subjected to mechanical refinement with full electrostatics, employing Charmm parameters in Sybyl. After refinement, it was observed that coordinates of the octet core were not significantly altered and that backbone parameters within the loop domains were within acceptable energetic limits. First, the structure is very compact, nearly spherical, with the three loop regions and the 5' "GT tail" comprising the surface of the tetrad core. Based upon this structure, it appears likely that interaction with cellular macromolecules will be heavily dominated by the structures of these surface loops. In that regard, it may be inappropriate to think of such interactions as "tetrad binding." The inclusion of G-tetrads in such a structure may not be important as a recognition element per se, but instead provides a latticework upon which an orderly loop array is positioned. Further, although the loop regions do not appear to be under mechanical stress, they are short enough so that they possess very configurational freedom. Because of those severe length constraints, we have found that all feasible loop models display a distinct "rabbit ears" structure, wherein the two base planes of the loop region are unstacked, and point outward from the center of the octet core. Such rigid, unstacked, single strand loop character is very distinctive as compared to other known folded nucleic acid structures. Therefore, varying the sequence or chemical structure of these loops, one at a time, is necessary to determine if bonding interactions between these loops and cellular macromolecules are important to the observed anti-HIV activity. The structures described above possess a single G-octet core, which is known to be the minimum structure required for nucleation of tetrad formation. Therefore, when paired with the observed short loop size, the intramolecular tetrad structure proposed for 1100-15 is best described as meta-stable, relative to other more robust tetrads which have been described in the literature. An increase of the core from 2 to 3 stacked tetrads, or an increase in the length of flexibility of one or more loops would be expected to increase the thermodynamic and/or kinetic stability of this structure significantly. Thus, the observed anti-HIV activity can be improved by sequence modification which enhances the stability of the underlying tetrad latticework. Finally, it has been observed that 1100-15 and homologues display profound resistance to cellular nucleases. One interesting aspect of the proposed structure is that, even in the loop domains, phosphodiester linkages (red) are generally buried from interaction with large solutes, such as a nuclease. The structure analysis proposed defines local phosphodiester backbone structure at low resolution. When paired with explicit biochemical analysis of phosphodiester cleavage rate, it is possible to define sites for selective introduction of backbone modification in 1100-15 homologies, for the purpose of extending the biological half life in vivo. The gel electrophoresis data described above suggests that 1100-15 spends every little time as a random coil at 25° C, under native salt conditions. Although the gel data rules out intermolecular associations, the data do not constrain the oligomer to any particular folded monomeric structure. Oligonucleotide folding in 1100-15 has been studied employing a combination of high resolution NMR and methods. Stable formation of a discrete octet core, mediated by tight binding of a single monovalent ion is crucial to the model described above. Given that G-Nl imino protons give rise to sharp, characteristic H NMR signals in such a structure, focus has been on the potassium ion dependence and temperature dependence of 1100-15 folding, as assessed by H NMR at 500mHz. For these measurements, 1100-15 has been synthesized at 15uM scale employing fast deblocking "Expedite" chemistry on a Milligen synthesizer. Subsequent to purification by denaturing anion-exchange chromatography in base (lOmM LiOH, 0.2 to 0.7M NaCl), oligomer purity was confirmed by denaturing gel electrophoresis (7M urea, 65° C). For NMR, the oligomer was desalted and transferred into 20mM LiCl adjusted to pH 6.0, which minimizes folding to form tetrads. Oligonucleotide strand concentration was held constant at 2.7 mM. MMR was measured in H^, employing a Redfield pulse sequence to saturate the water resonance, as described previously, Dittrich et al., Biochemistry, 1994, in press. In Figure 16 a KCl titatation is displayed. At 300° K, in the absence of added K + , imino proton signals cannot be resolved in the 10-12 ppm region. Subsequent to addition of KCl, substantial narrowing of imino signals is obtained, saturating at an added KCl concentration of 3mM, which is very close to one added K + equivalent per octet. Above 4 mM it can be seen that at least two classes of imino resonance can be detected in the 10-12 ppm range with roughly equal intensity: a broad envelope from 10-11 ppm, upon which several sharp resonances are superimposed in the 11-11.5 ppm region. By analogy with chemical shifts of other G tetrad structures, we tentatively ascribe the sharp imino signals to the 8 Hoogsteen H bonds of the core octet. The broad envelope is ascribed to the G and T imino resonances contributed by the loop and 5' terminal domains. Consistent with published tetrad NMR data, a broad envelop of signal was detected at 9 ppm, which most likely results from unusually show exchange of guanosine N2 protons engaged in Hoogsteen pairing. In order to better distinguish the two classes of iminoΗ signal and, additionally, to investigate the gross stability characteristics of the folded 1100-15 structure, thermal melting analysis, at 2.7 mM in strands, 6mM KCl, 20mM LiCl, pH 6.0 over the range from 300° K to 345° K has been performed. Substantial line narrowing of "Hoogsteen" imino proton signals is seen at 310° K, which appears to be accompanied by broadening of the poorly resolved imino envelope at 10.7 ppm. This narrowing plateaus above 310° K, giving rise to 7-8 well-resolved imino protons at 320° K. By reference to the NMR behavior of the oxytricia tetrad and other tetrad structures, the formation of 7 to 8 narrow,well-resolved imino resonances at elevated temperatures strongly suggests that in the presence of one bound K + ion per octet equivalent, 1100-15 has folded into a discreet tetrad structure, stabilized by the 8 Hoogsteen H-bonds of the presumed octet. In the range from 330 to 340° K, the imino proton spectrum undergoes an abrupt transition, which is likely to be representative of cooperative unfolding of the octet. Stability of this kind, accompanied by apparently high thermal cooperativity is very striking indeed, and is generally indicative of a single, well-defined folded oligonucleotide structure. The origin of the shallow temperature dependence of the spectral parameters, leading to enhanced ~ H resolution at 320° K, remains to be determined. It is likely to have resulted from weak intermolecular association which occur in the millimolar strand concentration range. This interpretation is born out by preliminary analysis of spectral parameters as a function of strand concentration (not shown). Independent of interpretation, the data suggests that high quality NMR data may be obtained for exchangeable and non-exchangeable 1100-15 protons at 35° C, 20 mM, LiCl, 6mM KCl and 2mM in strand equivalents. EXAMPLE 11 INHIBITION OF HCMV ACTIVITY Several different oligonucleotides reduced HCMV titers in tissue culture. Each of the oligonucleotides contained a different percentage of guanosine residues and a different number of total nucleotides in the polymer. The results of this assay are depicted in Table 9. All oligonucleotides were capable of reducing viral titer in culture including G101-50 which contained only 53% G residues (16 out of 30 total nucleotides). In Table 9, the length and percent guanosine nucleotides is indicated for each oligonucleotide tested. TABLE 9 Oligonucleotide Inhibition of HCMV Activity Viral Yield in plaque forming units (PFU) oligonucleotide (%G) In NIH3T3 cells chronically infected with FMLV, oligonucleotides (Figure 1) were capable of inhibiting virus production. However, oligonucleotides controls in this experiment were capable of inhibiting virus production in culture. EXAMPLE 12 IN VITRO ENZYMATIC ASSAYS Culture media containing FMLV reverse transcriptase (RT) was mixed with various concentrations of 1100-51 or 1100-12, the phosphodiester backbone of 1100-51 having been modified to a phosphorothioate backbone. Reverse transcriptase was measured as described in Example 7. Figure 4 shows that both oligonucleotides were capable of inhibiting the RT enzyme. Inhibitory concentrations for 50% reduction in RT activity was between 0.5 to 1 μM for 1100-51 and less than 0.5 uM for 1100-12. The 1100-51 (FMLV2ap), attenuated full length transcription directed by either the T7 or T3 polymerases (Figure 5a). As can be seen in Figure 1, full length transcripts directed by the T7 promoter would be 131 bases long while full length transcripts directed by the T3 promoter would be 171 bases long (position of the Dde I site relative to the mRNA start site). The sequence isomer of 1100-51 (1100-01 = FMLV2p), designed parallel to the target strand was also capable of significantly inhibiting transcription from the T7 promoter (Figure 5b). However, only the anti-parallel triple helix forming oligonucleotide FMLV2ap inhibited via attenuation of transcription as can be seen in the build up of a truncated transcript in the reaction mix (Figure 5C). The truncated transcript analyzed in Figure 5C was approximately 63 bases long and matched the predicted size fragment when p275A was used as a template (T7 promoter). G101-50 (53% G) inhibited T7, but not T3 directed, transcription by a mechanism other than attenuation (Figure 5A) since no truncated transcripts were observed when this oligonucleotide was used alone. 1100-11 (26% G) increased the level of specific transcripts directed by the T7 promoter (Figure 4b). In experiments designed to monitor inhibition of transcription initiation of the HSV-1 IE 175 promoter, using oligonucleotides, both specific and control G-Rich oligonucleotides were capable of inhibiting eukaryotic transcription when a HeLa cell extract system was used. The oligonucleotides used were B133-54; B133-55 and B107-51 as specific inhibitors via potential triple helix mechanism of action and G101-50 and 1100-11 as the low G-content control oligonucleotides. The present invention demonstrates anti-viral activity in tissue culture assays for several G-Rich oligonucleotides against HSV-2, HIV-1, HCMV and FMLV. In addition, G-Rich oHgonucleotides specifically inhibited the bacterial RNA polymerase enzymes T7 and T3, the FMLV and HIV-1 reverse transcriptase enzyme and eukaryotic RNA polymerase. All patents and publications mentioned in this specification are indicative of the levels of those skilled in the art to which the invention pertains. All patents and publications are herein incorporated by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference. One skilled in the art will readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned as well as those inherent therein. The oligonucleotides, compounds, methods, procedures and techniques described herein are presently representative of preferred embodiments, are intended to be exemplary and are not intended as limitations on the scope. Changes therein and other uses will occur to those skilled in the art which are encompassed within the spirit of the invention or defined by the scope of the appended claims. SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: Rando, Robert F. Fennewald, Susan Zendegui, Joseph G. Ojwang, Joshua 0. Hogan, Michael E. (ii) TITLE OF INVENTION: Anti-Viral Guanosine-Rich Oligonucleotides (ϋi) NUMBER OF SEQUENCES: 52 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: Fulbright & Jaworski (B) STREET: 1301 McKinney, Suite 5100 (C) CITY: Houston (D) STATE: Texas (E) COUNTRY: U.S.A. (F) ZIP: 77010-3095 (V) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentln Release #1.0, Version #1.25 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: US (B) FILING DATE: 28-OCT-1993 (C) CLASSIFICATION: (vii) PRIOR APPLICATION DATA: (A) APPLICATION NUMBER: US 08/053,027 (B) FILING DATE: 23-APR-1993 (viii) ATTORNEY/AGENT INFORMATION: (A) NAME: Paul, Thomas D. (B) REGISTRATION NUMBER: 32,714 (C) REFERENCE/DOCKET NUMBER: D-5574-CIP (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: 713/651-5151 (B) TELEFAX: 713/651-5246 (C) TELEX: 762829 (2) INFORMATION FOR SEQ ID NO:l: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 38 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 38 (D) OTHER INFORMATION: /note= "Amine moity attached to 3' end" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l: TGGGTGGGGT GGGGTGGGGG GGTGTGGGGT GTGGGGTG 38 ( 2 ) INFORMATION FOR SEQ ID NO : 2 : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 38 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: GTGGGGTGTG GGGTGTGGGG GGGTGGGGTG GGGTGGGT 38 (2) INFORMATION FOR SEQ ID NO:3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:3 GGGTGGGTGG GTGGGTGG 18 (2) INFORMATION FOR SEQ ID NO:4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 38 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: GGTGGTGGGG GGGGGTGGGG TGGTGGTGGG GGTGTTGG 38 (2) INFORMATION FOR SEQ ID NO:5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 36 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:5: GTGGTGGTGG TGTTGGTGGT GGTTTGGGGG GTGGGG 36 (2) INFORMATION FOR SEQ ID NO:6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 36 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: GTGGTTGGTG GTGGTGTGTG GGTTTGGGGT GGGGGG 36 (2) INFORMATION FOR SEQ ID NO:7: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 36 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 36 (D) OTHER INFORMATION: /note= "phosphorothioate backbone" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: GTGGTGGTGG TGTTGGTGGT GGTTTGGGGG GTGGGG 36 (2) INFORMATION FOR SEQ ID NO:8: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 36 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 36 (D) OTHER INFORMATION: /note= "phosphorothioate backbone" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: GTGGTTGGTG GTGGTGTGTG GGTTTGGGGT GGGGGG 36 (2) INFORMATION FOR SEQ ID NO:9: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 31 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: GGTGGGGTGG TGGTGGTTGG GGGGGGGGGG T 31 (2) INFORMATION FOR SEQ ID NO:10: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10: GGTGGTTGGG GGGTGGGGGG G 21 (2) INFORMATION FOR SEQ ID NO:11: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11: GGGTGGGGTG GTGGGTGGGG G 21 (2) INFORMATION FOR SEQ ID NO:12: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 30 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12: GGTGGGTGGT TTGTGTGGTT GGTGGGTTTT 30 (2) INFORMATION FOR SEQ ID NO:13: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 31 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13: GGGGGGGGGG TGTGGGGGGG GGTTGTGGTG G 31 (2) INFORMATION FOR SEQ ID NO:14: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 27 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14: GGTGGGTGGG TTGGGGGGTG GGTGGGG 27 (2) INFORMATION FOR SEQ ID NO:15: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 29 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15: TGGGGTTTGG GTGGGGGGTT GGGTGGTTG 29 (2) INFORMATION FOR SEQ ID NO:16: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: iinear (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:16: GGGTGGTGGT GTTGGTGTTG TGTG 24 (2) INFORMATION FOR SEQ ID NO:17: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17: GGTGGGGGGG TTGGTGTGTT TG 22 (2) INFORMATION FOR SEQ ID NO:18: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:18: GTGTGGGGGG GTGGGGTGGG GTGGGT 26 (2) INFORMATION FOR SEQ ID NO:19: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19: GGGTGGGTGG GTGGGTGGGT GGGTGG 26 (2) INFORMATION FOR SEQ ID NO:20: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 26 (D) OTHER INFORMATION: /note= "Amine moity attached to 3' end" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20: GTTGGGGGTT GTTGGTGGGG TGGTGG 26 (2) INFORMATION FOR SEQ ID NO:21: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 45 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 45 (D) OTHER INFORMATION: /note= "Amine moity attached to 3' end" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21: TGGTGGGTGT GTGGGGGGTG TTGGGGGTTG TTGGTGGGGT GGTGG 45 (2) INFORMATION FOR SEQ ID NO:22: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 45 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 45 (D) OTHER INFORMATION: /note= "cholesterol moity attached to 3' end" (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:22: TGGTGGGTGT GTGGGGGGTG TTGGGGGTTG TTGGTGGGGT GGTGG 45 (2) INFORMATION FOR SEQ ID NO:23: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 45 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 45 (D) OTHER INFORMATION: /note= "cholesterol moity attached to 3' end" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23: GTGGTGGGTG GGTGGGTGGT GGGTGGTGGT TGTGGGTGGG TGGTG 45 (2) INFORMATION FOR SEQ ID NO:24: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 45 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) ( ix) FEATURE : (A) NAME/KEY: misc_feature (B) LOCATION: 45 (D) OTHER INFORMATION: /note= "Amine moity attached to 3' end" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24: GTGGTGGGTG GGTGGGTGGT GGGTGGTGGT TGTGGGTGGG TGGTG 45 (2) INFORMATION FOR SEQ ID NO:25: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 26 (D) OTHER INFORMATION: /note= "cholesterol moity attached to 3' end" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25: GTTGGGGGTT GTTGGTGGGG TGGTGG 26 (2) INFORMATION FOR SEQ ID NO:26: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 45 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 45 (D) OTHER INFORMATION: /note= "Amine moity attached to 3' end" (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:26: GATCCATGTC AGTGACACTG CGTAGATCCG ATGATCCAGT CGATG 45 (2) INFORMATION FOR SEQ ID NO:27: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 26 (D) OTHER INFORMATION: /note= "phosphorothioate backbone and amine moity attached to backbone" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27: GTTGGGGGTT GTTGGTGGGG TGGTGG 26 (2) INFORMATION FOR SEQ ID NO:28: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28: GGTGGTGGGG TGGTTGTTGG GGGTTG 26 (2) INFORMATION FOR SEQ ID NO:29: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 45 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29: GGTGGTGGGG TGGTTGTTGG GGGTTGTTGG GGGTGTGTGG GTGGT 45 (2) INFORMATION FOR SEQ ID NO:30: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30: GGGTGGTTGG GTGGTTGG 18 (2) INFORMATION FOR SEQ ID NO:31: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 18 (D) OTHER INFORMATION: /note= "Amine moity attached to 3' end" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31: GGGTGGGTGG GTGGGTGG 18 (2) INFORMATION FOR SEQ ID NO:32: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 18 (D) OTHER INFORMATION: /note= "Amine moity attached to 3' end and phosphothioate backbone" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32: GGGTGGGTGG GTGGGTGG 18 (2) INFORMATION FOR SEQ ID NO:33: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 17 (D) OTHER INFORMATION: /note= "Amine moity attached to 3' end" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33: GTGGTGGGTG GGTGGGT 17 (2) INFORMATION FOR SEQ ID NO:34: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 27 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 27 (D) OTHER INFORMATION: /note= "Amine moity attached to 3 ' end" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34: GTGGTGGGTG GGTGGGTGGT GGGTGGT 27 (2) INFORMATION FOR SEQ ID NO:35: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 37 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 37 (D) OTHER INFORMATION: /note= "Amine moity attached to 3' end" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35: GTGGTGGGTG GGTGGGTGGT GGGTGGTGGT TGTGGGT 37 (2) INFORMATION FOR SEQ ID NO:36: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 16 (D) OTHER INFORMATION: /note= "Amine moity attached to 3' end" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36: TTGTGGGTGG GTGGTG 16 (2) INFORMATION FOR SEQ ID NO:37: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 29 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 29 (D) OTHER INFORMATION: /note= "Amine moity attached to 3' end" (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:37: TGGTGGGTGG TGGTTGTGGG TGGGTGGTG 29 (2) INFORMATION FOR SEQ ID NO:38: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 38 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 38 (D) OTHER INFORMATION: /note= "Amine moity attached to 3' end" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38: GTGGGTGGGT GGTGGGTGGT GGTTGTGGGT GGGTGGTG 38 (2) INFORMATION FOR SEQ ID NO:39: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 45 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 45 (D) OTHER INFORMATION: /note= "phosphorothioate backbone and amine moity attached to 3' end" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39: GTGGTGGGTG GGTGGGTGGT GGGTGGTGGT TGTGGGTGGG TGGTG 45 (2) INFORMATION FOR SEQ ID NO:40: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 18 (D) OTHER INFORMATION: /note= "Amine moity attached to 3' end" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40: GATCCATGTC AGTGACAC 18 (2) INFORMATION FOR SEQ ID NO:41: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 18 (D) OTHER INFORMATION: /note= "Amine moity attached to 3' end and phosphorothioate backbone" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41: GATCCATGTC AGTGACAC 18 (2) INFORMATION FOR SEQ ID NO:42: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 18 (D) OTHER INFORMATION: /note= "Amine moity attached to 3' end" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42: CCCCCCCCCC CCCCCCCC 18 (2) INFORMATION FOR SEQ ID NO:43: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 18 (D) OTHER INFORMATION: /note= "Amine moity attached to 3' end and phosphorothioate backbone" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43: CCCCCCCCCC CCCCCCCC 18 (2) INFORMATION FOR SEQ ID NO:44: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 47 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44: TTCATTTGGG AAACCCTTGG AACCTGACTG ACTGGCCGTC GTTTTAC 47 (2) INFORMATION FOR SEQ ID NO:45: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 15 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45: GTAAAACGAC GGCCA 15 (2) INFORMATION FOR SEQ ID NO:46: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46: GTGGTGGGTG GGTGGGG 17 (2) INFORMATION FOR SEQ ID NO:47: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:47: GTGGTGGGTG GGTGGG 16 (2) INFORMATION FOR SEQ ID NO:48: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48: TGGTGGGTGG GTGGGT 16 (2) INFORMATION FOR SEQ ID NO:49: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 13 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49: GTGGTGGGTG GGT 13 (2) INFORMATION FOR SEQ ID NO:50: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 9 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50: GTGGTGGGT (2) INFORMATION FOR SEQ ID NO:51: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:51: GTGGGTGGGT GGGT 14 (2) INFORMATION FOR SEQ ID NO:52: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52: GTGGGTGGGT 10

Claims: WHAT IS CLAIMED IS: 1. A method of treating a pathophysiological state caused by a virus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human or other animal, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide forms guanosine tetrads and wherein said tetrads form a stabilized three dimensional structure. 2. A method of treating a pathophysiological state caused by a virus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human or other animal, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains at least two runs of at least two guanosine bases. 3. A method of treating a pathophysiological state caused by a virus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human or other animal, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains a high percentage of guanosine bases. 4. The method of claims 1, 2 or 3, wherein said virus is selected from the group consisting of herpes simplex virus, human papilloma virus, Epstein Barr virus, human immunodeficiency virus, adenovirus, respiratory syncytial virus, hepatitis B virus and human cytomegalo virus. 5. The method of claims 1, 2 or 3, wherein said oligonucleotide is selected from the group of sequences consisting of SEQ ID Nos. 1-52. 6. A method of treating a pathophysiological state caused by a herpes simplex virus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide forms guanosine tetrads and wherein said tetrads form a stabilized three dimensional structure. 7. A method of treating a pathophysiological state caused by a herpes simplex virus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains at least two runs of at least two guanosine bases. 8. A method of treating a pathophysiological state caused by a herpes simplex virus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains a high percentage of guanosine bases. 9. The method of claims 6, 7 or 8, wherein said oligonucleotide is selected from the group of sequences consisting of SEQ ID Nos. 5-11. 10. A method of treating a pathophysiological state caused by human immunodeficiency virus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide forms guanosine tetrads and wherein said tetrads form a stabilized three dimensional structure. 11. A method of treating a pathophysiological state caused by human immunodeficiency virus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains at least two runs of at least two guanosine bases. 12. A method of treating a pathophysiological state caused by human immunodeficiency virus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains a high percentage of guanosine bases. 13. The method of claims 10, 11 or 12, wherein said oligonucleotide is selected from the group of sequences consisting of SEQ ID Nos. 1-4, 18-39, and 46-52. 14. A method of inhibiting HIV-1 activity comprising contacting HIV-1 infected cells with an HIV-1 Reverse Transcriptase inhibiting amount of an HrV-1 guanosine/thymidine oligonucleotide. 15. A method of inhibiting an HIV- 1 infection comprising contacting HIV-1 infected cells with an HIV-1 p24 production inhibiting amount of an HIV-1 guanosine/thymidine oligonucleotide. 16. The method of claims 10, 11, 12, 14 or 15, wherein the amount of the oligonucleotide inhibits syncytia formation. 17. A method of inhibiting the interaction of the outer envelope glycoprotein gpl20 of HIV-1, with CD4 cell surface glycoprotein, comprising binding said gpl20 with a sufficient dose of phosphorothioate substituted oligonucleotide to inhibit the interaction, wherein said substituted oligonucleotide forms guanosine tetrads and wherein said tetrads form a stabilized three dimensional structure. 18. A method of inhibiting the interaction of the outer envelope glycoprotein gpl20 of HIV-1, with CD4 cell surface glycoprotein, comprising binding said gpl20 with a sufficient dose of phosphorothioate substituted oligonucleotide to inhibit the interaction, wherein said oligonucleotide contains at least two runs of at least two guanosine bases. 19. A composition for inhibiting HIV-1 activity comprising, a guanosine-rich oligonucleotide having at least about 40% guanosine nucleotides; and a nucleotide sequence substantially non-homologous and substantially non-complementary to the HIV-1 genome. 20. A composition for inhibiting HIV-1 activity comprising an oligonucleotide capable of forming guanosine tetrads and wherein said tetrads form a stabilized three dimensional structure. 21. A composition for inhibiting HIV-1 activity comprising an oligonucleotide having at least two runs of at least two guanosine bases. 22. A method of treating a pathophysiological state caused by human papilloma virus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide forms guanosine tetrads and wherein said tetrads form a stabilized three dimensional structure. 23. A method of treating a pathophysiological state caused by human papilloma virus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains at least two runs of at least two guanosine bases. 24. A method of treating a pathophysiological state caused by human papilloma virus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains a high percentage of guanosine bases. 25. A method of treating a pathophysiological state caused by human cytomegalovirus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide forms guanosine tetrads and wherein said tetrads form a stabilized three dimensional structure. 26. A method of treating a pathophysiological state caused by human cytomegalovirus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains at least two runs of at least two guanosine bases. 27. A method of treating a pathophysiological state caused by human cytomegalovirus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains a high percentage of guanosine bases. 28. The method of claims 25, 26 or 27, wherein said oligonucleotide is selected from the group of sequences consisting of SEQ ID Nos. 12-17. 29. A method of treating a pathophysiological state caused by hepatitis B virus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide forms guanosine tetrads and wherein said tetrads form a stabilized three dimensional structure. 30. A method of treating a pathophysiological state caused by hepatitis B virus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains at least two runs of at least two guanosine bases. 31. A method of treating a pathophysiological state caused by hepatitis B virus, comprising the step of administering a pharmacological dose of an oligonucleotide to a human, said dose being sufficient to inhibit the replication of said virus, wherein said oligonucleotide contains a high percentage of guanosine bases. 32. The method of one of the claims of 1-3, 6-8, 10-12, 17, 18, 21-27 or 29-31, wherein said percentage of guanosine bases in said oligonucleotide is at least about 40%. 33. The method of one of the claims of 1-3, 6-8, 10-12, 17, 18, 21-27 or 29-31, wherein said oligonucleotide is capped at the 3' terminus with a modifier selected from the group consisting of polyamine, poly-L-lysine and cholesterol. 34. The method of claim 33, wherein the cap is propanolamine. 35. The method of one of the claims of 1-3, 6-8, 10-12, 17, 18, 21-27 or 29-31, wherein said oligonucleotide is capped at the 5' terminus with a modifier selected from the group consisting of polyamine, poly-L-lysine and cholesterol. 36. The method of claim 35, wherein the cap is propanolamine. 37. The method of one of the claims of 1-3, 6-8, 10-12, 17, 18, 21-27 or 29-31, wherein said oligonucleotide has a modified phosphodiester linkage. 38. The method of claim 37, wherein said oligonucleotide is modified to contain a phosphorothioate linkage. 39. A guanosine-rich oligonucleotide, wherein said oligonucleotide exhibits anti-viral activity. 40. The oligonucleotide of claim 39, wherein said oligonucleotide contains at least about 40% guanosine bases. 41. The oligonucleotide of claim 39 selected from the group consisting of SEQ. ID. NOs. 1 to 52. 42. The oligonucleotide of claim 43 or 44 having a backbone at least partially substituted with a phosphorothioate moiety. 43. The oligonucleotide of claim 39, 40 or 41, wherein said oligonucleotide has an end terminal capped with a modifier selected from the group consisting of polyamine, poly-L-lysine and cholesterol. 44. The oligonucleotide of any of claims 39, 40 or 41, wherein said oligonucleotide has an end terminal capped with a propanolamine modifier.

Inventors: FENNEWALD SUSAN, HOGAN MICHAEL E, OJWANG JOSHUA O, RANDO ROBERT F, ZENDEGUI JOSEPH G, FENNEWALD, SUSAN, HOGAN, MICHAEL, E., OJWANG, JOSHUA, O., RANDO, ROBERT, F., ZENDEGUI, JOSEPH, G.
Applicants: BAYLOR COLLEGE MEDICINE, FENNEWALD SUSAN, HOGAN MICHAEL E, OJWANG JOSHUA O, RANDO ROBERT F, TRIPLEX PHARMA CORP, ZENDEGUI JOSEPH G, BAYLOR COLLEGE OF MEDICINE, FENNEWALD, SUSAN, HOGAN, MICHAEL, E., OJWANG, JOSHUA, O., RANDO, ROBERT, F., TRIPLEX PHARMACEUTICAL CORPORATION, ZENDEGUI, JOSEPH, G.
Application year: 1994
Main classification: A61K31
IPC codes: A61K31, A61K38, A61P31, C07H21, C12N15